Lec 2: Introduction to TE:

Scaffold
Shah Rukh Abbas, PhD
3-1-2021

https://www.ibiology.org/bioengineering/tissue-engineering/
Organs are made up of tissues.
• Products of the synthesis can be tissues or
organs.
• Almost all organs are essentially made up of
three types of tissues: epithelial, basement
membrane and stroma (connective tissue).
• Members of the Tissue Triad
– EPITHELIA 100% cells. No matrix. No blood vessels.
– BASEMENT MEMBRANE No cells. 100% matrix. No
blood vessels.
– STROMA (CONNECTIVE TISSUE) Cells. Matrix. Blood
vessels.

• When excised from an organ, the epithelia are

regenerated spontaneously. Examples: the
epidermis in skin, the myelin sheath in nerves.
• Likewise, the basement membrane regenerates
spontaneously on the stroma.
• However, the stroma does not regenerate
spontaneously. Examples: dermis in skin,
endoneurium in nerves.
 Which tissues in the triad do not regenerate
spontaneously?
• When excised from an organ, the epithelia
are regenerated spontaneously. Examples:
the epidermis in skin, the myelin sheath in
nerves.
• Likewise, the basement membrane
regenerates spontaneously on the stroma.
• However, the stroma does not regenerate
spontaneously. Examples: dermis in skin,
endoneurium in nerves.
• Epithelia and basement membrane (BM) are
synthesized from remaining epithelial cells.
• The stroma is not synthesized from remaining
stromal cells. Instead, these cells induce closure
of the injury by contraction and synthesis of scar.
• Therefore, the central question in organ
synthesis is how to synthesize the stroma.
• Once the stroma has been induced to
synthesize, epithelial cells can spontaneously
synthesize both epithelia and BM over it
(“sequential” synthesis).
 3 Tools of Tissue Engineering

• Cells
– Living part of tissue
– Produces protein and provides function of cells
– Gives tissue reparative properties
• Scaffold
– Provides structural support and shape to construct
– Provides place for cell attachment and growth
– Usually biodegradable and biocompatible
• Cell Signaling
– Signals that tell the cell what to do
– Proteins or Mechanical Stimulation
1. Scaffold
 What do we want in a scaffold?
1. Biocompatible
2. Biodegradable
3. Chemical and Mechanical Properties
4. Proper architecture
 Requirements of Scaffold
• Biocompatibility
– It must be biocompatible; cells must adhere, function normally, and
migrate onto the surface and eventually through the scaffold and begin to
proliferate before laying down new matrix.
– After implantation, the scaffold or tissue engineered construct must elicit a
negligible immune reaction in order to prevent it causing such a severe
inflammatory response that it might reduce healing or cause rejection by
the body
• Biodegradability
– The objective of tissue engineering is to allow the body’s own cells, over
time, to eventually replace the implanted scaffold or tissue engineered
construct.
– The scaffold must therefore be biodegradable so as to allow cells to
produce their own extracellular matrix.
– In order to allow degradation to occur in tandem with tissue formation, an
inflammatory response combined with controlled infusion of cells such as
macrophages is required
• Mechanical properties
– Ideally, the scaffold should have mechanical properties consistent with the
anatomical site into which it is to be implanted and, from a practical
perspective, it must be strong enough to allow surgical handling during
implantation
– Producing scaffolds with adequate mechanical properties is one of the
great challenges in attempting to engineer bone or cartilage. For
particularly cardiovascular and orthopaedic tissues, the implanted scaffold
must have sufficient mechanical integrity to function from the time of
implantation to the completion of the remodelling process.
– A further challenge is that healing rates vary with age; for example, in
young individuals, fractures normally heal to the point of weight-bearing
in about six weeks, with complete mechanical integrity not returning until
approximately one year after fracture, but in the elderly the rate of repair
slows down
– balance between mechanical properties and porous architecture sufficient
to allow cell infiltration and vascularization is key to the success of any
scaffold
• Scaffold architecture
– Scaffolds should have an interconnected pore structure and high porosity
to ensure cellular penetration and adequate diffusion of nutrients to cells
within the construct and to the extra-cellular matrix formed by these cells.
– It is required to allow diffusion of waste products out of the scaffold, and
the products of scaffold degradation should be able to exit the body
without interference with other organs and surrounding tissues.
– Cells primarily interact with scaffolds via chemical groups (ligands) on the
material surface. Scaffolds synthesized from natural extracellular materials
(e.g. collagen) naturally possess these ligands in the form of Arg-Gly-Asp
(RGD) binding sequences (Fig. 2), whereas scaffolds made from synthetic
materials may require deliberate incorporation of these ligands through,
for example, protein adsorption. The ligand density is influenced by the
specific surface area, i.e. the available surface within a pore to which cells
can adhere. This depends on the mean pore size in the scaffold.
The pores thus need to be large enough to allow cells to migrate
into the structure, where they eventually become bound to the
ligands within the scaffold, but small enough to establish a
sufficiently high specific surface, leading to a minimal ligand density
to allow efficient binding of a critical number of cells to the scaffold
• Manufacturing technology
In order for a particular scaffold or tissue engineered construct to
become clinically and commercially viable, it should be cost
effective, and it should be possible to scale-up from making one at
a time in a research laboratory to small batch production.

Another key factor is determining how a product will be delivered

and made available to the clinician. This will determine how either
the scaffold or the tissue engineered construct will be stored.
Clinicians typically prefer off-the shelf availability without the
requirement for extra surgical procedures in order to harvest cells
prior to a number of weeks of in vitro culture before implantation.
However, for some tissue types, this is not possible and in vitro
engineering prior to implantation is required.
 Polymers in TE
• Synthetic
Natural
– Made
Derived
byfrom
controlled
ECM process
• ++/-preprogrammed
Range of biological responses
• + Generally
+/- Range of degradation
• biocompatible
Established production protocol
• +Biological
degradation
mechanism
• - not made, purified
 Architecture
• Pore size
– Average diameter of pores
• Porosity
– Porosity volume/total volume
• Interconnectivity
• Porogeneration (melt molding)
 Methods of Cell Delivery
• Cell Adhesion (solid/dry scaffolds)

• Cell Encapsulation
– Polymer solution to solid
Extracellular matrix (ECM)
Analogy between scaffold and ECM